内蒙古农业科技
內矇古農業科技
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INNER MONGOLIA AGRICULTURAL SCIENCE AND TECHNOLOGY
2014年
5期
102-105
,共4页
菊花%白晨%李晓东%包南帝娜%郭永青
菊花%白晨%李曉東%包南帝娜%郭永青
국화%백신%리효동%포남제나%곽영청
抑制消减杂交%植物%基因差异表达
抑製消減雜交%植物%基因差異錶達
억제소감잡교%식물%기인차이표체
Suppression subtractive hybridization (SSH)%Plant%gene differential expression
在植物的不同生长、发育、繁殖阶段和不同类型细胞中,基因的表达各不相同。近几年发展起来的研究基因差异表达的方法很多,抑制消减杂交(Suppression Subtractive Hybridization)是其中一项基于转录水平,结合消减杂交与 PCR技术的分离差异表达基因的方法,其原理可概括为消减同源序列,富集差异序列。该技术报道于1996年,在一个循环过程中分离差异表达基因尤其是低丰度表达的基因,虽然存在一些缺点,但以其较高的灵敏性、容易操作、假阳性率低、重复性好等优点,在分离及其克隆差异表达基因研究方面得到了广泛的应用。近几年,此技术在植物抗病基因的诱导表达方面已得到了应用,目前大多数研究主要集中在文库构建及筛选等方面。文章对抑制性消减杂交(SSH)技术的原理、优缺点及其在植物抗病性研究机制中应用进展进行了综述。
在植物的不同生長、髮育、繁殖階段和不同類型細胞中,基因的錶達各不相同。近幾年髮展起來的研究基因差異錶達的方法很多,抑製消減雜交(Suppression Subtractive Hybridization)是其中一項基于轉錄水平,結閤消減雜交與 PCR技術的分離差異錶達基因的方法,其原理可概括為消減同源序列,富集差異序列。該技術報道于1996年,在一箇循環過程中分離差異錶達基因尤其是低豐度錶達的基因,雖然存在一些缺點,但以其較高的靈敏性、容易操作、假暘性率低、重複性好等優點,在分離及其剋隆差異錶達基因研究方麵得到瞭廣汎的應用。近幾年,此技術在植物抗病基因的誘導錶達方麵已得到瞭應用,目前大多數研究主要集中在文庫構建及篩選等方麵。文章對抑製性消減雜交(SSH)技術的原理、優缺點及其在植物抗病性研究機製中應用進展進行瞭綜述。
재식물적불동생장、발육、번식계단화불동류형세포중,기인적표체각불상동。근궤년발전기래적연구기인차이표체적방법흔다,억제소감잡교(Suppression Subtractive Hybridization)시기중일항기우전록수평,결합소감잡교여 PCR기술적분리차이표체기인적방법,기원리가개괄위소감동원서렬,부집차이서렬。해기술보도우1996년,재일개순배과정중분리차이표체기인우기시저봉도표체적기인,수연존재일사결점,단이기교고적령민성、용역조작、가양성솔저、중복성호등우점,재분리급기극륭차이표체기인연구방면득도료엄범적응용。근궤년,차기술재식물항병기인적유도표체방면이득도료응용,목전대다수연구주요집중재문고구건급사선등방면。문장대억제성소감잡교(SSH)기술적원리、우결점급기재식물항병성연구궤제중응용진전진행료종술。
The genes of plants were differently expressed in various phases of growth、development and multiplication and in different cells. In recent years, numerous methods used to investigate the gene differential expression were developed. Suppression Subtractive Hybridization (SSH), developed in 1996, is a technique with high efficiency for isolation differently expressed genes at transcriptional level, combining subtractive hybridization with PCR, and its principle can be summarized as eliminating homologous sequences and enriching different sequences. SSH is able to isolate the differently expressed genes in one cycle, especially for genes expressed with low abundance. The features that high specificity, simple operation, low false positive rate and good repeatability make it widely applied in the differential expression genes separation despite of some defects. In recent years, SSH has been applied in studies on induced expression of plant disease resistant genes, with most focus on cDNA library construction and screen. The principle, main advantages and defects, and application progress in mechanism study on plants disease resistance were reviewed in this paper.